Fig. 1. Serum activity of the osteoclast-specific enzyme TRAP-5b and serum levels of RANKL and OPG, key regulators involved in osteoclast differentiation.

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Fig. 1. Serum activity of the osteoclast-specific enzyme TRAP-5b and serum levels of RANKL and OPG, key regulators involved in osteoclast differentiation in 42 stable renal transplant patients. Values are given as median, 25–75th percentile (bars) and minimum/maximum (whiskers), *P < 0.05. From: Impact of sirolimus, tacrolimus and mycophenolate mofetil on osteoclastogenesis—implications for post-transplantation bone disease Nephrol Dial Transplant. 2011;26(12):4115-4123. doi:10.1093/ndt/gfr214 Nephrol Dial Transplant | © The Author 2011. Published by Oxford University Press on behalf of ERA-EDTA. All rights reserved. For Permissions, please e-mail: journals.permissions@oup.com

Fig. 2. Multinucleated osteoclast precursor cells and their TRAP expression after 10 days of culture in the presence of M-CSF and RANKL. Values are given as mean ± SD, *P < 0.05 versus control. From: Impact of sirolimus, tacrolimus and mycophenolate mofetil on osteoclastogenesis—implications for post-transplantation bone disease Nephrol Dial Transplant. 2011;26(12):4115-4123. doi:10.1093/ndt/gfr214 Nephrol Dial Transplant | © The Author 2011. Published by Oxford University Press on behalf of ERA-EDTA. All rights reserved. For Permissions, please e-mail: journals.permissions@oup.com

Fig. 3. Osteoclast activity was detected after prolonged culture for 28 days in the presence of RANKL and M-CSF on hydroxyapatite-coated wells. After von Kossa staining, the area of resorbed mineral was quantified by its bright appearance in the transmitted light. Values are given as mean ± SD, *P < 0.05 versus control. From: Impact of sirolimus, tacrolimus and mycophenolate mofetil on osteoclastogenesis—implications for post-transplantation bone disease Nephrol Dial Transplant. 2011;26(12):4115-4123. doi:10.1093/ndt/gfr214 Nephrol Dial Transplant | © The Author 2011. Published by Oxford University Press on behalf of ERA-EDTA. All rights reserved. For Permissions, please e-mail: journals.permissions@oup.com

Fig. 4. Real-time PCR analysis in osteoclast precursors following 8 days of culture in the presence of recombinant M-CSF/RANKL. Target gene expression was normalized for 1 million copies ribosomal RNA (18S). Results are given as mean ± SD and represent five independent experiments of different donors. The mean value of the control groups was assigned to 1, *P < 0.05 versus control. From: Impact of sirolimus, tacrolimus and mycophenolate mofetil on osteoclastogenesis—implications for post-transplantation bone disease Nephrol Dial Transplant. 2011;26(12):4115-4123. doi:10.1093/ndt/gfr214 Nephrol Dial Transplant | © The Author 2011. Published by Oxford University Press on behalf of ERA-EDTA. All rights reserved. For Permissions, please e-mail: journals.permissions@oup.com

Fig. 5. Real-time PCR analysis in MSC Fig. 5. Real-time PCR analysis in MSC. Target gene expression was normalized for 1 million copies ribosomal RNA (18S). Each column represents the mean of independent culture experiments of a total of four different donors. The mean value of the control groups was assigned to 1. From: Impact of sirolimus, tacrolimus and mycophenolate mofetil on osteoclastogenesis—implications for post-transplantation bone disease Nephrol Dial Transplant. 2011;26(12):4115-4123. doi:10.1093/ndt/gfr214 Nephrol Dial Transplant | © The Author 2011. Published by Oxford University Press on behalf of ERA-EDTA. All rights reserved. For Permissions, please e-mail: journals.permissions@oup.com

Fig. 6. Effect of SRL on the viability of mononuclear cells/osteoclast precursors at different time points after isolation as detected by Annexin V-FITC and propidium iodide staining with additional CD14 PE labelling and subsequent FACS analysis. As depicted in the upper panel, addition of SRL led to some induction of apoptosis. However, apoptosis promoting actions of SRL could largely be prevented by M-CSF and RANKL co-administration. Omitting M-CSF and RANKL from the culture medium was more pro-apoptotic than the effect of SRL treatment. SRL, M-CSF and RANKL primarily influenced mononuclear cell survival as depicted by the fraction of CD14-positive cells in the various treatment groups and time points. From: Impact of sirolimus, tacrolimus and mycophenolate mofetil on osteoclastogenesis—implications for post-transplantation bone disease Nephrol Dial Transplant. 2011;26(12):4115-4123. doi:10.1093/ndt/gfr214 Nephrol Dial Transplant | © The Author 2011. Published by Oxford University Press on behalf of ERA-EDTA. All rights reserved. For Permissions, please e-mail: journals.permissions@oup.com

Fig. 7. ( A ) Overview of osteoclastogenesis from haematopoietic precursor cells into mature osteoclasts. RANKL and RANK are essential for osteoclast maturation and activation. OPG can bind and antagonize RANKL, thereby inhibiting osteoclastogenesis (schema adapted from W.J. Boyle [ 1 ]). ( B ) Gives an overview about the key in vitro and in vivo findings how SRL, CI and MMF may affect osteoclast maturation at different stages. From: Impact of sirolimus, tacrolimus and mycophenolate mofetil on osteoclastogenesis—implications for post-transplantation bone disease Nephrol Dial Transplant. 2011;26(12):4115-4123. doi:10.1093/ndt/gfr214 Nephrol Dial Transplant | © The Author 2011. Published by Oxford University Press on behalf of ERA-EDTA. All rights reserved. For Permissions, please e-mail: journals.permissions@oup.com